Orchestration of adhesion signalling by the mechanosensors talin and vinculin.

通过机械传感器 talin 和 vinculin 协调粘附信号。

• 批准号: BB/P000681/1
• 负责人: Christoph Ballestrem
• 金额: $ 55.45万
• 依托单位: University of Manchester
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2016
• 资助国家: 英国
• 起止时间: 2016 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FP000681%2F1
• 关键词: Orchestration; adhesion; signalling; mechanosensors; talin

Cells continuously sense and produce their surrounding environment, which consists of fibrillar material the cells can attach to and is called extracellular matrix (ECM). Cell-ECM communication is particularly important during development or regeneration processes that require specific cellular responses to changing environments. Cellular responses comprise changes in motile behaviour (e.g. closing of wounds), contractility (e.g. functioning of the cardiovascular system) but also active remodelling of their ECM for the purpose of formation of new functional tissue. Many studies have focused on how cells sense their environment, but we are still far from understanding the mechanisms how cells perceive environmental signals and how they are translated into signals within cells that promote specific cellular responses.The environment of cells alters enormously during development, normal ageing, injury and certain diseases. For example, the mechanical properties of the ECM is thought to influence tumour progression and increased breast matrix stiffness is associated with poor survival. Stiffening of ECM also causes cardiovascular malfunctioning. Intriguingly cells contribute to the production of specific matrix on one hand but also respond to this produced environment on the other hand. Therefore, understanding how cells sense and produce their ECM environment is critically important if we want to get a step closer to treating the roots of diseases and promote regeneration.Cells can feel or sense their environment by exerting forces on it and probing its deformation. To transmit forces, they 'grab' neighbouring structures using surface proteins, which are called integrins. These integrins not only bind to the environment of the cells but also connect to a skeleton inside the cells. This link is not direct but is regulated by components that couple or uncouple the two. We published a number of manuscripts showing that two of these coupling proteins, called talin and vinculin, are central to sensing of environmental changes. They are particularly important to measure the stiffness of their environment, they control cell migration, as well as cell growth and differentiation. In this proposal we also present important pilot data demonstrating that vinculin is critical for ECM remodelling. However how they do this is still unclear. In order to investigate how these proteins regulate the response to their environment, and to what extent they are involved in telling cells how to behave, the two laboratories in the prestigious Cell-Matrix Centre at the University of Manchester will team up and combine their long-standing expertise with the field of integrins signalling and cell-matrix interactions. The proposed research aims to to (i) understand the role of mechanical signals in the activation of talin and vinculin, (ii) how this activation helps vinculin and talin to associate with a large number other proteins that serve to exert specific signals (e.g. cell migration or cell growth) (iii) how vinculin with the newly found association of another protein called tensin is contributing to the formation and remodelling of ECM environment. To reach our goals, we will not only use cutting edge microscopy, biochemisty and molecular biology techniques but also a newly generated intracellular system whereby we can target proteins to specific compartments (mitochondria) in the cells which enable us to visualise and probe molecular interactions and behaviour under defined conditions. Our results will be combined into a model that outlines and potentially predicts how cells interpret and remodel their environment. Ultimately, the knowledge gained may lead to important changes in how we currently envisage environmental changes and their contribution to diseases. This may also lead to changes in treatment of patients, and it might thus, for example, contribute to improvements in disease prevention and in regeneration processes.

细胞不断感知并产生周围环境，该环境由细胞可以附着的纤维材料组成，称为细胞外基质 (ECM)。细胞-ECM 通讯在发育或再生过程中尤其重要，因为发育或再生过程需要特定的细胞对不断变化的环境做出反应。细胞反应包括运动行为（例如伤口闭合）、收缩性（例如心血管系统功能）的变化，还包括为了形成新的功能组织而主动重塑其 ECM。许多研究都集中在细胞如何感知环境上，但我们还远未了解细胞感知环境信号的机制以及它们如何转化为细胞内促进特定细胞反应的信号。细胞的环境在发育过程中发生巨大变化，正常情况下衰老、受伤和某些疾病。例如，ECM 的机械特性被认为会影响肿瘤的进展，而乳腺基质硬度的增加与生存率较低有关。 ECM 硬化还会导致心血管功能障碍。有趣的是，细胞一方面有助于特定基质的产生，另一方面也对这种产生的环境做出反应。因此，如果我们想进一步治疗疾病的根源并促进再生，了解细胞如何感知和产生其 ECM 环境至关重要。细胞可以通过对其施加力并探测其变形来感知或感知其环境。为了传递力，它们使用表面蛋白（称为整合素）“抓住”邻近的结构。这些整合素不仅与细胞环境结合，还与细胞内的骨架连接。该链接不是直接的，而是由将两者耦合或分离的组件调节。我们发表的许多手稿表明，其中两种耦合蛋白（称为talin和vinculin）对于感知环境变化至关重要。它们对于测量环境的硬度特别重要，它们控制细胞迁移以及细胞生长和分化。在此提案中，我们还提供了重要的试点数据，证明纽蛋白对于 ECM 重塑至关重要。然而他们如何做到这一点仍不清楚。为了研究这些蛋白质如何调节对其环境的反应，以及它们在多大程度上参与告诉细胞如何行为，曼彻斯特大学著名的细胞基质中心的两个实验室将合作并结合他们的长期研究成果。 - 整合素信号传导和细胞-基质相互作用领域的长期专业知识。拟议的研究旨在（i）了解机械信号在踝蛋白和纽蛋白激活中的作用，（ii）这种激活如何帮助纽蛋白和踝蛋白与大量其他蛋白质结合，这些蛋白质用于施加特定信号（例如细胞） (iii) 纽蛋白与新发现的另一种称为张力蛋白的蛋白质的结合如何促进 ECM 环境的形成和重塑。为了实现我们的目标，我们不仅将使用尖端的显微镜、生物化学和分子生物学技术，而且还将使用新生成的细胞内系统，通过该系统，我们可以将蛋白质靶向细胞中的特定区室（线粒体），从而使我们能够可视化和探测分子相互作用和在规定条件下的行为。我们的结果将被整合到一个模型中，该模型概述并可能预测细胞如何解释和重塑其环境。最终，所获得的知识可能会导致我们目前对环境变化及其对疾病的影响的看法发生重大变化。这也可能导致患者治疗的改变，因此可能有助于改善疾病预防和再生过程。

项目成果

Distinct focal adhesion protein modules control different aspects of mechanotransduction.

不同的粘着斑蛋白模块控制力转导的不同方面。

• DOI: http://dx.10.1242/jcs.195362
• 发表时间: 2017
• 期刊: Journal of cell science
• 影响因子: 4
• 作者: Stutchbury B

Mechanotransduction at the cell-matrix interface.

细胞-基质界面的力转导。

• DOI: http://dx.10.1016/j.semcdb.2017.07.027
• 发表时间: 2017
• 期刊: Seminars in cell & developmental biology
• 影响因子: 7.3
• 作者: Jansen KA

Talin gets SHANKed in the fight for integrin activation.

塔林在整合素激活的斗争中被刺伤。

• DOI: http://dx.10.1038/ncb3501
• 发表时间: 2017
• 期刊: Nature cell biology
• 影响因子: 21.3
• 作者: Atherton P

Low-intensity pulsed ultrasound promotes cell motility through vinculin-controlled Rac1 GTPase activity.

低强度脉冲超声通过纽蛋白控制的 Rac1 GTPase 活性促进细胞运动。

• DOI: http://dx.10.1242/jcs.192781
• 发表时间: 2017
• 期刊: Journal of cell science
• 影响因子: 4
• 作者: Atherton P

Light-Induced Molecular Adsorption of Proteins Using the PRIMO System for Micro-Patterning to Study Cell Responses to Extracellular Matrix Proteins.

使用 PRIMO 微图案系统进行光诱导蛋白质分子吸附，研究细胞对细胞外基质蛋白质的反应。

• DOI: http://dx.10.3791/60092
• 发表时间: 2019
• 期刊: JoVE
• 作者: Melero C